FIGURE 7.

Gremlin via Notch pathway induces EMT in cultured human tubular epithelial cells. Cells (HK2 cell line) were preincubated with DAPT (30 nM) before stimulation with Gremlin (10 ng/ml). Changes in EMT-related markers were assessed by confocal microscopy (A) or western blot (B). Please, note that the pictures of e-cadherin, α-sma and their corresponding loading control (GAPDH) are the same images as shown in the Figure 2B but, in this case, Gremlin + DAPT point is included in the fourth lane. Results are expressed as mean ± SEM of five independent experiments. All changes induced by Gremlin were prevented by Notch inhibition. Figure (A) shows a representative experiment out of two performed by confocal microscopy and (B) shows representative images of western blot and quantification. (C) Gene expression levels were evaluated 24 h after Gremlin stimulation. Total cell RNA was isolated to assess mRNA levels by RT-qPCR. Data are expressed as mean ± SEM of six independent experiments. ^∗p < 0.05 vs. control; ^#p < 0.05 vs. Gremlin.